Arabidopsis thealiana AT3G56230 AT4G18650

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Arabidopsis thealianaAT3G56230AT4G18650

• What are the functions of them?
• Are they important for seed development?

Spring 2004 Mariko Onozawa
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• What is AT3G56230?
• About 1200bps.
• 5UTR.
• Three SALK lines.
• This protein containing BTB/POZ domain.

• What is AT4G18650?
• About 1400bps.
• 5 and 3UTR.
• No SALK line.
• This protein is related to transcription
• factor and has weak similarity to
• TGAGG-sequence specific DNA-
• binding protein(TGA-2.1).

RV
FV
FV
RV
3

• mRNA
• Find the pattern of mRNA concentration
• RT-PCR (SRB)
• Genechip database
• Gene
• Find KOs of my gene
• Madison lines
• SALK lines
• Phenotype
• Study a phenotype of plants which containing the
  KO
• Check genotypes of the plants
  (SALK)

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• RT-PCR result using SRB (AT3G56230)
• Genechip data
• RT-PCR result says that the gene which is
• homologous toAT3G56230 is expressed at all
• stages and organs of SRB.
• Genechip data says that AT3G56230
• is expressed only at Mature green seed,
• Seedling (3DAI), Roots and stem.

A border between presence and absence
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Is there any KO of my gene in Madison lines?

• AT3G56230
• I couldnt find any KO of AT3G56230.
• The insertion site is upstream of the gene.
  (-594bp)
• AT4G18650
• I couldnt find any KO of AT4G18650.
• The insertion site is downstream of the gene.

R19
FV
RV
R15
FV
RV
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Checking phenotypes of the plants
?WT, T-DNA
?WT
?T-DNA
?WT, T-DNA
?T-DNA
?WT
Some seedlings dont have any band.
Some samples dont have any band.
nothing
Only tubulins bands
Nothing
Only tubilans bands
T-DNA,homo 1,4 WT, homo 2,3,5,6

1 2 3 4 5 6 WC
1 2 3 4 5 6 WC
smears
?WT, T-DNA
smears
1 2 3 4 5 6WC
1 2 3 4 5 6WC
?WT T-DNA
?WT, T-DNA
?WT, T-DNA
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I couldnt find any significant difference
between WT and mutant plants.
T-DNA
Wild Type
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What should be done next to study the functions?
Get more plants which have a insertion in the
genes. Observe and study the knockout
plants. Study with other knockout lines.
The final goal is to answer these questions.
What is the function of the protein? What is
the structure of the protein? How it is
regulated?
What we did in this class is only the beginning
of the research.